Isolation and characterization of porcine natural killer (NK) cells.

The morphologic and biological properties of porcine cells mediating natural killer (NK) activity were determined. In a previous study, we demonstrated that lymphocytes from the peripheral blood of pigs greater than 1 week of age possessed NK activity to K562 tumor cells and that lymphocytes from the blood and spleen of pigs greater than 1 day of age were able to mediate natural cytotoxicity against parainfluenza-3 (PI3) virus-infected Vero cells (Yang and Schultz, 1986a). Discontinuous density gradients were used to enrich NK cells. NK cytotoxicity was mainly present in high-density Percoll fractions (50 to 55% and 55 to 60%); little or no NK activity was present in lower density fractions. The NK cell enriched lymphocytes responded to the mitogens PHA, ConA and PWM. NK cells were sensitive to the suppressive effect of corticosteroid, but Protein A did not affect NK activity. The amount of cytotoxicity directly corresponded to the degree of binding that occurred between the NK enriched lymphocyte population and the target cells. Cytochemical and morphological studies demonstrated that these bond cells which are believed to be responsible for the NK activities, were mainly small to medium lymphocytes lacking azurophilic cytoplasmic granules. These findings were confirmed by ultrastructural studies of effector and "target-binding" cells. The results of the present study suggested that the cells mediating NK activity in pigs have the morphological and density characteristics of small and medium sized lymphocytes; findings that differ from those described for NK cells in human and other animal species.

Techniques for venous blood sampling from the white pekin duck.

Common sites for blood sampling in the pekin duck and other avian species include the basilic vein, jugular vein, superficial plantar metatarsal vein, heart, and occipital sinus. The use of each of these sites is described and/or illustrated. In the present study using the pekin duck, the superficial plantar metatarsal vein proved most satisfactory for collecting repeated samples (every 5 minutes for 30 minutes) with minimal trauma to the duck.

Baseline hematologic, endocrine, and clinical chemistry values in ducks and roosters.

Venous blood samples were collected at 3-day intervals for a total of six samples from each of five adult male pekin ducks and five adult Ross roosters. Twenty biochemical, six hematologic, and three endocrine determinations were performed on each blood or serum sample collected. The data obtained provide reference values for future studies of avian species and illustrate the utility of an automated clinical chemistry analyzer in assessing multiple serum biochemistry values in small sample volumes obtained from birds.

Ameloblastic fibrosarcoma in a bull.

This report describes a malignant odontogenic neoplasm in a 7-year-old bull. The mass, involving the right mandible, was locally invasive and destructive. Histologically, it consisted of islands and cords of benign odontogenic epithelium, entrapped in a population of malignant mesenchymal cells. These morphological features are characteristic of ameloblastic fibrosarcoma in man, an odontogenic tumour not previously described in animals.

Clinical and clinicopathologic effects of large doses of raw linseed oil as compared to mineral oil in healthy horses.

The clinical and clinicopathologic effects of raw linseed oil and mineral oil were compared. In a crossover experimental design trial, 6 horses were given either raw linseed oil (2.5 mL/kg body weight) or mineral oil (10 mL/kg body weight), twice, 12 hours apart. Two weeks later, the horses received the opposite treatment. All horses given mineral oil or linseed oil developed nonformed feces by 24 hours of the first administration of oil. Horses treated with mineral oil had formed feces at 48 hours; horses treated with linseed oil developed normally formed feces at 96 to 108 hours. All horses treated with linseed oil had signs of depression and anorexia, and 3 had signs of mild colic. These signs were not observed in horses treated with mineral oil. Concentrations of serum glucose and bilirubin were significantly higher in horses treated with linseed oil when compared with horses treated with mineral oil.

Effect of intestinal resection on two juvenile horses with granulomatous enteritis.

Two horses were presented with lethargy, weight loss, anorexia, and swelling of the limbs and ventral body wall. One horse, a 12-month-old American Paso Fino colt, also had acute abdominal pain. The other horse, a seven-month-old Tennessee Walking Horse (TWH) filly passed diarrheic stools during the initial examination. Each horse had low serum protein, neutropenia, and a normal packed cell volume (3.2 g/dl, 1300 cells/ul, and 38%, respectively, for the colt, and 2.4 g/dl, 696 cells/ul, and 44%, respectively for the filly). After intravenously administering plasma, the colt's PCV dropped to 23%, and the filly's dropped to 30%. During exploratory surgery, 3.5 and 2.0 meters of thickened terminal small intestine were removed from the colt and filly respectively, and a jejunocecostomy performed. The results of histologic examination of resected intestine were consistent with a diagnosis of equine granulomatous enteritis (EGE). Both horses showed clinical improvement within two days after surgery. The colt developed a neutrophilia (20,500 cells/ul) within 24 hours of surgery. Serum protein concentrations remained stable and gradually elevated to normal or near normal values of 7.0 g/dl (colt) and 5.8 g/dl (filly) by two weeks. The colt was killed four months after surgery because of signs of abdominal pain. Postmortem examination revealed a small intestinal volvulus associated with an adhesion. The TWH filly remains clinically normal 13 months after surgery.

Urine cortisol:creatinine ratio in healthy and sick cats.

Urine cortisol:creatinine ratios (UCCR) were determined from single urine samples obtained by cystocentesis in 47 cats allotted into 2 groups: 31 healthy cats and 16 sick, hospitalized cats with assorted clinical illnesses. The mean (+/- standard deviation) UCCR for healthy cats was 5.9 +/- 7.0 (median, 3.2; range, 0.6 to 27.8). Age or gonadal status had no significant effect on the magnitude of UCCR within this group. However, sick cats had significantly higher UCCR (P = .002) when compared with healthy cats. The mean UCCR for sick cats was 19.6 +/- 19.2 (median, 14.8; range, 1.7 to 75.1). This report establishes a reference range for UCCR in 31 normal cats and provides evidence that health status affects UCCR in cats.

Effects of castration on peritoneal fluid in the horse.

Twenty-four clinically normal horses were castrated by routine methods. Peritoneal fluid was collected prior to castration and at 1, 3, 5, and 7 days postcastration. Peritoneal fluid was collected on days 9 and 11 if nucleated cell (NC) counts were still markedly elevated on day 7. Peritonitis, defined as NC counts greater than 10,000/microliters, was evident in 15 horses following castration. Mean NC counts peaked on day 5 but were less than 10,000/microliters for 74% of the horses by day 7, and 90% of the horses by day 9. One horse had a NC count greater than 60,000/microliters on day 11 when sampling ended. Postcastration peritoneal fluid was obviously blood-tinged in 21 horses. Peak RBC counts occurred on day 3 but markedly decreased by day 5. Elevated peritoneal RBC counts correlated well with elevated NC counts (P less than 0.001). Horses with peritonitis tended to have fever (P less than 0.05). Other clinical signs of peritonitis were not apparent.

Cerebrospinal fluid composition of cattle with endotoxin-induced mastitis treated with isotonic (0.9%) or hypertonic (7.5%) sodium chloride.

This study examined the safety of intravenous hypertonic saline in cattle with experimental gram-negative endotoxemia. Cerebrospinal fluid (CSF) composition was examined in five control cows and eight treated cows 24 hours after the intramammary infusion of 1 mg of endotoxin. Four of the endotoxin challenged cows were treated intravenously with isotonic (0.9%) sodium chloride and four cows were treated intravenously with hypertonic (7.5%) sodium chloride. Decreased CSF osmolality, and sodium and alpha globulin concentrations and increased CSF concentrations of beta globulin were observed in both endotoxin-challenged saline-treated groups. No CSF compositional differences were observed between endotoxin-challenged cows receiving isotonic or hypertonic saline. Although no cytologic or biochemical evidence of salt poisoning was observed in cows receiving hypertonic saline, significant changes were observed in the CSF composition of both endotoxin-infused saline-treated groups.

Characterization of equine alkaline phosphatase isoenzymes based on their electrophoretic mobility by polyacrylamide gel disc electrophoresis.

Alkaline phosphatase isoenzymes of equine tissues, peritoneal fluid, and serum were characterized by their electrophoretic mobilities, using polyacrylamide gel disc electrophoresis. The alkaline phosphatase isoenzymes in liver, kidney, spleen, small intestine, placenta, bone, small colon, and large colon tissue samples were extracted and separated by electrophoresis. The resulting isoenzyme mobilities and spectrophotometric scans were evaluated for their tissue specificity and for their possible use in determining the tissue contribution of alkaline phosphatase to serum and peritoneal fluid. The sensitivity of the tissue alkaline phosphatase to heating at 56 C was also determined. The isoenzymes of all the tissue extracts, except small intestine and large colon, were distinguishable from one another by their specific electrophoretic mobilities and patterns and by their sensitivity to heating at 56 C. The mobilities and patterns of small intestine and large colon isoenzyme extracts were not different enough to allow making a differentiation, but the heat resistance of small intestine did distinguish it from large colon. The tissue sources of alkaline phosphatase in the serum of healthy adult horses, pregnant mares, and normal foals and in the peritoneal fluid of healthy horses were determined by characterization of the mobilities of their isoenzymes.

Cerebrospinal fluid cytology in canine neurologic disease.

Samples of cerebrospinal fluid (CSF) of 93 dogs with neurologic diseases were examined by cytomorphologic technique, and the changes in the CSF were correlated with histopathologic examinations of the central nervous system (CNS). It was concluded that CSF examination is a significant aid in obtaining a neurologic diagnosis and that good correlation exists between the CSF changes and the pathologic changes in the CNS. The CSF examination allows making a diagnosis of encephalitis and differentiation between viral and other causes (although in mycotic infection the cell membrane preparation can be used to identify the cause directly), could allow making differentiation between congenital malformations and congenital degenerative disease, and helps in identifying physical spinal cord damage, differentiating it from muscular, neurogenic, or functional disorders clinically presented as spinal ataxia. The CSF cytologic examination can indicate the presence of hemorrhage in the CNS. There is not enough experience available in the diagnosis of brain tumors by means of CSF examination; however, in dogs with lymphosarcoma in the CNS, CSF cytologic changes can be diagnostic.

Evaluation of sodium carboxymethylcellulose for prevention of experimentally induced abdominal adhesions in ponies.

Twelve ponies were used to evaluate the reliability of an abdominal adhesion model and the efficacy of intraperitoneal infusion of sodium carboxymethylcellulose in preventing abdominal adhesions. A celiotomy was performed on each of the 12 ponies and the serosa of the distal portion of the jejunum was abraded with a dry gauze sponge at 5 locations. In addition to the serosal damage, a single 2-0 chromic gut suture was placed through the seromuscular layer of the jejunum in the center of the abraded area. After closure of the celiotomy, a 1% solution of sodium carboxymethylcellulose (7 ml/kg of body weight) was infused into the peritoneal cavity of 6 ponies. The other ponies served as untreated controls. All ponies were euthanatized 14 days after surgery. All ponies in the control group had abdominal adhesions at the time of necropsy. Four of the 6 ponies in the treatment group were free of adhesions. There was a significant (P less than 0.0001) difference in the total number of adhesions between the 2 groups.

Enhanced platelet reactivity in heartworm-infected dogs.

Platelet number, mean platelet volume, and platelet function were evaluated in 34 clinically normal dogs and 28 heartworm-infected (HWI) dogs. Mean platelet numbers for dogs of the HWI group was not significantly lower than those for dogs of the control group (214,000 vs 254,000 cells/microliter); however, 6 (21%) HWI dogs had platelet numbers less than 150,000/microliter, compared with only 2 (6%) heartworm-negative dogs. The mean platelet volume was not significantly different (7.8 vs 7.7 fl) between the 2 groups of dogs. Mean platelet aggregation responses to intermediate and low concentrations of collagen (3.0 and 1.5 micrograms) and to high, intermediate, and low concentrations of ADP (25, 10, and 5 microM) were greater in dogs of the HWI group. Mean platelet 14C-serotonin release was also greater in HWI dogs in response to high concentration of ADP (25 microM) and to intermediate concentration of collagen (3.0 micrograms).

Evaluation of antithrombin-III activity as a coindicator of disseminated intravascular coagulation in cats with induced feline infectious peritonitis virus infection.

Six adult specific-pathogen-free cats were inoculated intraperitoneally with a cell culture-adapted strain of feline infectious peritonitis virus. Plasma samples were evaluated for antithrombin-III (AT-III) activities at post-inoculation days (PID) 0, 4, and 11 and at termination on PID 16 (1 cat) or 21 (5 cats). Other hemostatic values evaluated were activated partial thromboplastin times, prothrombin times, thrombin times, fibrinogen, platelet counts, and fibrin/fibrinogen degradation products. Antithrombin-III activity remained within normal or above normal range (89 to 246%) in all cats, with the exception of one cat on PID 4 (AT-III, 70%). Mean baseline AT-III activity for 6 cats at PID 0 was 123%. Mean AT-III activity on PID 4, 11, and 16 or 21 was 98, 162, and 130%, respectively. On PID 4 and 16 or 21, results of coagulation screening tests indicated that all cats had disseminated intravascular coagulation. Histologically, cats also had severe fibrinonecrotizing thrombovasculitis.

Serum gamma-glutamyl transpeptidase activity in healthy cats and cats with induced hepatic disease.

Activities of serum gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) were determined in healthy cats and in cats before and after treatment: common bile duct ligation, carbon tetrachloride administration, sham surgery, or anesthesia only. Significant (P less than 0.01) increases in serum GGT, ALP, and ALT occurred in cats with ligated bile ducts. Significant (P less than 0.01) increases in serum ALT occurred in carbon tetrachloride-treated cats. Increases of serum GGT, ALP, or ALT were not observed in cats subjected to sham surgery or anesthesia only compared with these cats' baseline values and values in healthy cats. Tissue GGT activity was measured in liver, renal cortex, jejunal mucosa, and bile ducts. There was a 1.5-fold increase in GGT activity in livers of cats with ligated bile ducts, compared with that in livers of healthy cats.

Application of an enzyme-multiplied immunoassay technique for determination of caffeine elimination kinetics as a test of liver function in clinically normal dogs.

A commercially available automated enzyme-multiplied immunoassay technique (EMIT) was used to determine serum caffeine concentration after oral and IV administrations of caffeine at dosage of 5 mg/kg of body weight to 12 clinically normal dogs. Dogs were allotted to 2 groups of 6 dogs each; 1 group initially received caffeine orally and the other received caffeine IV. After 72 hours, caffeine administration was repeated in all dogs in the alternate manner. Serum samples were obtained at multiple intervals over 24 hours to determine distribution and elimination kinetics. Analysis of the drug concentration-time data indicated IV elimination half-life (t1/2) of 6.39 +/- 1.87 hours, volume of distribution at steady state of 685.3 +/- 132.2 ml/kg, total body clearance of 1.31 +/- 0.38 ml/min/kg, absorption t1/2 of 1.02 +/- 0.68 hour, oral elimination t1/2 of 6.53 +/- 2.72 hours, lag time after oral administration of 0.0614 +/- 0.0661 hour, highest measured concentration of 5.29 +/- 1.17 micrograms/ml, time to peak concentration of 2.74 +/- 1.30 hours, and bioavailability of 99.4 +/- 19.4%. Data from 6 dogs best fit a 1-compartment open model and those from 6 other dogs best fit a 2-compartment open model. On the basis of data from the 6 dogs that best fit a 2-compartment model, t1/2 of distribution was 0.58 +/- 0.72 hour. Data for oral administration best fit a single absorption phase and a single elimination phase. The increased availability and simplicity of the EMIT offers an opportunity to study the application of caffeine elimination for clinical evaluation of dogs with liver disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of plasma and buffy coat ascorbic acid concentrations in dogs before and after a 24-hour fast.

Samples of blood were taken from 15 female and 15 male research laboratory Beagles before and after they were fasted for 24 hours. The mean buffy coat ascorbic acid concentration was significantly higher in dogs after they were fasted than that before they were fasted. In contrast, the mean plasma ascorbic acid concentration was significantly lower in dogs after they were fasted than that before they were fasted. The mean buffy coat ascorbic acid concentrations in blood samples of both fasted and nonfasted female Beagles was significantly greater than those of male Beagles, whereas the mean plasma concentrations of both fasted and nonfasted female Beagles was significantly lower than those of male Beagles. It was observed that whenever there was a decrease in plasma ascorbic acid concentration, there was an increase in buffy coat ascorbic acid concentration, regardless of fasting stress or sex difference.

Effects of treatment with aspirin or aspirin/dipyridamole combination in heartworm-negative, heartworm-infected, and embolized heartworm-infected dogs.

To determine the drug dose required to inhibit platelet reactivity by at least 50%, 2 drug regimens were evaluated in heartworm-negative, heartworm-infected, and heartworm-infected dogs embolized with dead heartworms. Aspirin, or a combination of aspirin and dipyridamole, were administered to 2 groups of Beagles (n = 5 each) for 5 to 9 days; a third group of 5 Beagles served as nontreated controls. For heartworm-negative dogs, mean (+/- SD) aspirin dosage that inhibited collagen-induced platelet reactivity by at least 50% was 6 (+/- 2) mg/kg of body weight given once daily. The aspirin/diphridamole combination dosage was 1 mg of each drug/kg given every 12 hours. All dogs (n = 15) were implanted with 7 adult heartworms each and remedicated (or not treated) beginning at 21 days after heartworm implantation. In heartworm-infected dogs, mean aspirin dosage required to inhibit collagen-induced platelet reactivity greater than or equal to 50% was 10 (+/- 6) mg/kg. Mean dosage of aspirin/dipyridamole combination was 1.6 +/- (0.5) mg of each drug/kg given every 12 hours. When platelet reactivity in response to collagen was determined to be inhibited by at least 50% in all medicated dogs, each dog (n = 15) was embolized with 7 dead adult heartworms to mimic heartworm adulticidal treatment. Platelet reactivity was monitored for 21 days after treatment, and drug dose was adjusted to maintain platelet inhibition by at least 50%. In embolized dogs, mean aspirin dosage was 17 (+/- 14) mg/kg given once daily. Mean dosage of the aspirin/dipyridamole combination was 2.8 (+/- 1.3) mg of each drug/kg given every 12 hours. All dogs (n = 15) were euthanatized 21 days after heartworm embolization. Each lung lobe was evaluated for severity of lesions and presence of organized or fibrinous thrombi. Lesion severity in the aspirin- and aspirin/dipyridamole-treated dogs was not significantly different from that in control dogs.

Effects of treatment with ticlopidine in heartworm-negative, heartworm-infected, and embolized heartworm-infected dogs.

Ticlopidine hydrochloride was evaluated for its effectiveness in inhibiting platelet aggregation and serotonin release in 5 laboratory Beagles before and after heartworm implantation with 7 adult Dirofilaria immitis, and after embolization with 7 dead heartworms to mimic what happens after heartworm adulticide treatment. Five other laboratory Beagles, similarly implanted and embolized with heartworms, were used as nonmedicated controls. During the heartworm-negative stage, the dosage of ticlopidine that inhibited adenosine diphosphate (ADP)-induced platelet aggregation in 5 dogs by at least 50% after 5 days of treatment was 62 mg/kg of body weight once a day. In the same dogs implanted with 7 adult heartworms 21 days previously, mean (+/- SD) ticlopidine dosage required to obtain similar results was 71 (+/- 13) mg/kg given once daily. During the 21 days after dead heartworms were implanted in heartworm-infected dogs, mean ticlopidine dosage was 108 (+/- 35) mg/kg (range, 62 to 150 mg/kg). Ticlopidine treatment was associated with increased platelet numbers in all 5 dogs during the heartworm-negative stage and in 4 of 5 dogs during the heartworm implantation and heartworm embolization stages. Mean platelet volume tended to decrease as platelet numbers increased. At necropsy, gross and histologic pulmonary lesions were less severe in ticlopidine-treated dogs than in nonmedicated control dogs.

Effects of dexamethasone and surgical hypotension on the stomach of dogs: clinical, endoscopic, and pathologic evaluations.

Effects of dexamethasone and surgical hypotension on the gastric mucosa of 15 dogs was evaluated by clinical, endoscopic, and pathologic studies. Dexamethasone produced acute reddening of the gastric mucosa with frank hemorrhage, digested blood, and possible gastric erosions, whereas dogs treated by surgical hypotension did not have marked changes in the gastric mucosa. Gastrointestinal disease observed in dogs with spontaneous spinal cord injury that were given dexamethasone and decompressive spinal surgery was not produced. The role of corticosteroids and surgical hypotension in the pathogenesis of gastric lesions and the potential for spinal cord injury to produce gastrointestinal disease is discussed.